Determination of tildipirosin residues in swine tissues by high performance liquid chromatography-tandem mass spectrometry
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摘要:目的
建立适用于猪组织中泰地罗新残留检测的高效液相色谱−串联质谱法。
方法猪组织(肌肉、肝脏、肾脏和皮脂)样品经0.1 mol·L–1磷酸二氢钾溶液提取，上清液采用HLB固相萃取柱浓缩净化，Phenomenex Luna Omega C18色谱柱梯度洗脱，采用电喷雾正离子扫描和多反应监测(MRM)模式对泰地罗新进行分析，采用基质匹配法对泰地罗新含量进行标准校正。
结果泰地罗新添加剂量在肌肉中25~2 400 ng·g–1、肝脏中25~2 500 ng·g–1、肾脏中25~2 000 ng·g–1和皮脂中25~1 600 ng·g–1的范围内，与特征离子峰面积呈良好的线性关系(r2 ≥ 0.99)。猪组织添加泰地罗新样品中肌肉、肝脏、肾脏和皮脂的检测限均为10 ng·g–1，定量限均为25 ng·g–1。对猪组织在定量限、1/2MRL、MRL、2MRL这4个剂量下进行泰地罗新的添加回收试验，各组织中的批内平均回收率为85.6%~105.0%，批内回收率相对标准偏差2.3%~9.5%，批间回收率相对标准偏差4.7%~7.6%。
结论该方法简单易行，灵敏度高且特异性强，可用于猪组织中泰地罗新残留量的分析测定。
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关键词:
- 猪组织 /
- 高效液相色谱–串联质谱 /
- 泰地罗新 /
- 残留
Abstract:ObjectiveTo establish a reliable HPLC-MS/MS (High performance liquid chromatography-tandem mass spectrometry) method suitable for determing tildipirosin residues in swine tissues.
MethodSwine tissue samples, including muscle, liver, kidney and skin-fat were extracted with 0.1 mol·L–1 KH2PO4 buffer solution. The supernatant fluids were enriched and purified using HLB solid-phase extraction column, and gradiently eluted by Phenomenex Luna Omega C18 liquid chromatography column. The analytes were then detected using triple-quadrupole mass spectrometry in positive electrospray ionization and multiple reaction monitoring (MRM) mode. The matrix-matched method was used to calibrate tildipirosin content.
ResultTildipirosin contents presented good linear relationships (r2>0.99) with characteristicion peak area in the ranges of 25–2 400 ng·g–1 in muscle, 25–2 500 ng·g–1 in liver, 25–2 000 ng·g–1 in kidney and 25–1 600 ng·g–1 in skin-fat. All the limits of detection and quantitation in muscle, liver, kidney and skin-fat samples added tildipirosin were 10 and 25 ng·g–1, respectively. The recovery experiments of tildipirosin in swine tissues were setted in four dosage levels, including limit of quantitation, 1/2 maximum residue limit (MRL), MRL and 2MRL, and the mean intra-batch recoveries of tildipirosin in all analytes ranged from 85.6% to 105.0%. The relative standard deviations of intra-batch and inter-batch recoveries were in the ranges of 2.3%–9.5% and 4.7%–7.6%, respectively.
ConclusionThe established method is simple, practicable, and with high sensitivity and specificity. It can be applied to determine tildipirosin residues in swine tissues.
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图 3 空白肌肉样品及添加25 ng·g–1泰地罗新的肌肉样品的多反应检测谱图
Figure 3. Multiple reaction monitoring chromatograms of blank muscle sample and muscle sample added 25 ng·g–1 tildipirosin
表 1 猪组织中泰地罗新检测的基质匹配标准曲线
Table 1 The martrix-matched standard curves of tildipirosin detected in swine tissues
组织样品 w (泰地罗新)1)/
(ng·g–1)标准曲线方程 2) r2 肌肉 25~2 400 y=109x+120 0.995 5 肝脏 25~2 500 y=124x−201 0.998 7 肾脏 25~2 000 y=55.1x+133 0.997 3 皮脂 25~1 600 y=50.8x+214 0.997 5 1）猪组织中泰地罗新的添加剂量范围；2）x：猪组织中泰地罗新的添加剂量，y：泰地罗新特征离子峰面积 
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表 2 猪组织中添加泰地罗新样品回收率及相对标准偏差
Table 2 Recoveries and relative standard deviations of swine tissue samples added tildipirosin
组织
样品w (泰地罗新)添加/
(ng·g–1)批内平均回收率/% 批内回收率相对标准偏差/% 批间平均
回收率/%批间回收率相对
标准偏差/%批次1 批次2 批次3 批次1 批次2 批次3 肌肉 25 101.0 96.6 96.3 6.6 7.3 5.3 97.8 6.4 600 93.3 100.0 97.6 8.5 4.3 8.1 97.1 7.4 1 200 97.5 95.5 97.9 9.5 5.6 6.6 97.0 7.1 2 400 99.0 98.5 99.6 4.7 7.9 6.3 99.1 6.1 肝脏 25 99.1 96.8 101.0 6.7 8.0 7.1 99.1 7.1 2 500 99.9 99.1 101.0 6.4 6.8 3.1 100.0 5.4 5 000 96.0 101.0 93.3 5.8 2.3 4.6 96.7 5.3 10 000 96.7 100.0 93.9 5.4 2.6 6.2 96.9 5.3 肾脏 25 101.0 98.4 101.0 4.3 8.9 6.1 100.0 6.4 5 000 92.3 96.1 97.2 6.0 4.9 2.9 95.2 5.0 10 000 85.6 91.5 93.8 2.5 4.5 4.5 90.3 5.5 20 000 93.3 90.0 96.1 4.9 3.3 3.7 93.1 4.7 皮脂 25 100.0 105.0 102.0 6.3 4.2 4.8 102.0 5.2 400 94.6 102.0 90.7 3.2 4.1 4.6 95.8 6.3 800 92.3 94.4 91.1 8.2 5.9 6.5 92.6 6.7 1 600 92.4 94.7 91.5 9.0 5.6 8.8 92.9 7.6
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